Gold nano-seeds (GNSs) (capital EF, Cyrillic=2-5 nm) were dotted in TiO(2) colloids and the horseradish peroxidase (HRP) was successfully immobilized on the as-made GNSs-TiO(2) nanocomposite by a convenient and effective method. The matrix integrates the merits of both GNSs and TiO(2), which provides a favorable microenvironment for the immobilization of HRP. The cyclic votammetric results demonstrated that the entrapped HRP achieves direct electron transfer at glassy carbon electrode (GCE). A pair of stable and quasi-reversible redox peaks with a small peak-to-peak separation of 43 mV was observed in phosphate buffer solution. The GNSs stabilized by TiO(2) colloids acted sufficiently as the conducting tunnel to promote the electron transfer. As a result, the electrochemical behaviors were improved in virtue of the synergic effect of TiO(2) and GNSs. The Nafion/HRP-GNSs-TiO(2)/GCE displayed an excellent and rapid electrocatalytic response to the reduction of H(2)O(2). The proposed biosensor exhibited a good linear response in the range from 4.1 x 10(-5) to 6.3 x 10(-4) mol L(-1), with a detection limit of 5.9 x 10(-6) mol L(-1) (at the ration of signal to noise, S/N=3). The apparent Michaelis-Menten constant was estimated to be 0.63 mmol L(-1). Furthermore, the biosensor possesses satisfactory stability and good reproducibility.
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